LOCUS       sequence-383873-        6535 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the M. jannaschii tRNATyr (MjtRNA) and GFP(47TAG) (aka 
            GFP151TAG).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6535)
  AUTHORS   Touchette MH, Van Vlack ER, Bai L, Kim J, Cognetta AB 3rd, Previti 
            ML, Backus KM, Martin DW, Cravatt BF, Seeliger JC
  TITLE     A Screen for Protein-Protein Interactions in Live Mycobacteria 
            Reveals a Functional Link between the Virulence-Associated Lipid 
            Transporter LprG and the Mycolyltransferase Antigen 85A.
  JOURNAL   ACS Infect Dis. 2017 May 12;3(5):336-348. doi: 
            10.1021/acsinfecdis.6b00179. Epub 2017 Mar 21.
  PUBMED    28276676
REFERENCE   2  (bases 1 to 6535)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acsinfecdis.6b00179"; journalName: "ACS Infect Dis"; date: 
            "2017-05-12- 12"; volume: "3"; issue: "5"; pages: "336-348"
COMMENT     Sequence Label: pSMT-MjtRNA-GFP(47TAG)
FEATURES             Location/Qualifiers
     source          1..6535
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      11..599
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     500..519
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     753..770
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    887..908
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(923..928,929..946,947..953)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    961..977
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     966..988
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     985..1001
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     985..1001
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(1443..1445,1446..2159)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDAT*GKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1485..1506)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1746..1765)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2093..2114
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2330..2346)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2331..2347)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
ORIGIN
        1 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
       61 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
      121 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
      181 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
      241 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
      301 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
      361 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
      421 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
      481 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
      541 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
      601 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
      661 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
      721 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
      781 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
      841 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      901 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      961 ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacc agatctggct
     1021 cgcaccgcgg tggcggccgc tctagacggt gaccacaacg cgcccgcttt gatcggggac
     1081 gtctgcggcc gaccatttac gggtcttgtt gtcgttggcg gtcatgggcc gaacatactc
     1141 acccggatcg gagggccgag gacaaggtcg aacgaggggc atgacccggt gcggggcttc
     1201 ttgcactcgg cataggcgag tgctaagaat aacgttggca ctcgcgaccg gtgagtgcta
     1261 ggtcgggacg gtgaggccag gcccgtcgtc gcagcgagtg gcagcgagga caacttgagc
     1321 cgtccgtcgc gggcactgcg cccggccagc gtaagtagcg gggttgccgt cacccggtga
     1381 cccccgtttc atccccgatc cggaggaatc acttcgcaat ggccaagaca attgcggatc
     1441 ccgtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     1501 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctagg
     1561 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     1621 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     1681 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     1741 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     1801 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     1861 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     1921 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     1981 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     2041 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     2101 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaac
     2161 atatggattt ggcgagcttc gtgcgtgttc ggtagcctgg catttaccga ccggcggtag
     2221 ttcagcaggg cagaacggcg gactctaaat ccgcatggcg ctggttcaaa tccggcccgc
     2281 cggaccagcg gaaatgaccc cagaggtgcg acctctgggg taagcttatc gataccgtcg
     2341 acctcgaggg ggggcccccg gggatcatcg agccgagaac gttatcgaag ttggtcatgt
     2401 gtaatcccct cgtttgaact ttggattaag cgtagataca cccttggaca agccagttgg
     2461 attcggagac aagcaaattc agccttaaaa agggcgaggc cctgcggtgg tggaacaccg
     2521 cagggcctct aaccgctcga cgcgctgcac caaccagccc gcgaacggct ggcagccagc
     2581 gtaaggcgcg gctcatcggg cggcgttcgc cacgatgtcc tgcacttcga gccaagcctc
     2641 gaacacctgc tggtgtgcac gactcacccg gttgttgaca ccgcgcgcgg ccgtgcgggc
     2701 tcggtggggc ggctctgtcg cccttgccag cgtgagtagc gcgtacctca cctcgcccaa
     2761 caggtcgcac acagccgatt cgtacgccat aaagccaggt gagcccacca gctccgtaag
     2821 ttcgggcgct gtgtggctcg tacccgcgca ttcaggcggc agggggtcta acgggtctaa
     2881 ggcggcgtgt acggccgcca cagcggctct cagcggcccg gaaacgtcct cgaaacgacg
     2941 catgtgttcc tcctggttgg tacaggtggt tgggggtgct cggctgtcgc tggtgttcca
     3001 ccaccagggc tcgacgggag agcgggggag tgtgcagttg tggggtggcc cctcagcgaa
     3061 atatctgact tggagctcgt gtcggaccat acaccggtga ttaatcgtgg tctactacca
     3121 agcgtgagcc acgtcgccga cgaatttgag cagctctggc tgccgtactg gccgctggca
     3181 agcgacgatc tgctcgaggg gatctaccgc caaagccgcg cgtcggccct aggccgccgg
     3241 tacatcgagg cgaacccaac agcgctggca aacctgctgg tcgtggacgt agaccatcca
     3301 gacgcagcgc tccgagcgct cagcgcccgg gggtcccatc cgctgcccaa cgcgatcgtg
     3361 ggcaatcgcg ccaacggcca cgcacacgca gtgtgggcac tcaacgcccc tgttccacgc
     3421 accgaatacg cgcggcgtaa gccgctcgca tacatggcgg cgtgcgccga aggccttcgg
     3481 cgcgccgtcg acggcgaccg cagttactca ggcctcatga ccaaaaaccc cggccacatc
     3541 gcctgggaaa cggaatggct ccactcagat ctctacacac tcagccacat cgaggccgag
     3601 ctcggcgcga acatgccacc gccgcgctgg cgtcagcaga ccacgtacaa agcggctccg
     3661 acgccgctag ggcggaattg cgcactgttc gattccgtca ggttgtgggc ctatcgtccc
     3721 gccctcatgc ggatctacct gccgacccgg aacgtggacg gactcggccg cgcgatctat
     3781 gccgagtgcc acgcgcgaaa cgccgaattc ccgtgcaacg acgtgtgtcc cggaccgcta
     3841 ccggacagcg aggtccgcgc catcgccaac agcatttggc gttggatcac aaccaagtcg
     3901 cgcatttggg cggacgggat cgtggtctac gaggccacac tcagtgcgcg ccagtcggcc
     3961 atctcgcgga agggcgcagc agcgcgcacg gcggcgagca cagttgcgcg gcgcgcaaag
     4021 tccgcgtcag ccatggaggc attgctatga gcgacggcta cagcgacggc tacaaccggc
     4081 agccgactgt ccgcaaaaag cggcgcgtga ccgccgccga aggcgctcga atcaccggac
     4141 tatccgaacg ccacgtcgtc cggctcgtgg cgcaggaacg cagcgagtgg ctcgccgagc
     4201 aggctgcacg ccgcgaacgc atccgcgcct atcacgacga cgagggccac tcttggccgc
     4261 aaacggccaa acatttcggg ctgcatctgg acaccgttaa gcgactcggc tatcgggcga
     4321 ggaaagagcg tgcggcagaa caggaagcgg ctcaaaaggc ccacaacgaa gccgacaatc
     4381 caccgctgtt ctaacgcaat tggggagcgg gtgtcgcggg ggttccgtgg ggggttccgt
     4441 tgcaacgggt cggacaggta aaagtcctgg tagacgctag ttttctggtt tgggccatgc
     4501 ctgtctcgtt gcgtgtttcg ttgcgcccgt tttgaatacc agccagacga gacggggttc
     4561 tacgaatctt ggtcgatacc aagccatttc cgctgaatat cggggagctc accgccagaa
     4621 tcggtggttg tggtgatgta cgtggcgaac tccgttgtag tgcctgtggt ggcatccgtg
     4681 gccactctcg ttgcacggtt cgttgtgccg ttacaggccc cgttgacagc tcaccgaacg
     4741 tagttaaaac atgctggtca aactaggttt accaacgata cgagtcagct catctagggc
     4801 cagttctagg cgttgttcgt tgcgcggttc gttgcgcatg tttcgtgtgg ttgctagatg
     4861 gctccgcaac cacacgcttc gaggttgagt gcttccagca cgggcgcgat ccagaagaac
     4921 ttcgtcgtgc gactgtcctc gttgatcctt gccgagctgg gatggaagcc cggccgaccc
     4981 accctggagg agatgatcga ggatgccagg gcctttcacg cccgccgctg ctgagcgtcc
     5041 gccgccgggc ccgcaccgcc gtcggccggc ccgctccggg ctcgcagcag cgggcttcgg
     5101 cgcgggcccg gggctcccgg gccgccgggc ggggctccgc ccggcggccg ccgggggccg
     5161 ggggcggcgc cgggcggccc ggggcgtcag gcgccggggg cggtgtccgg cggcccccag
     5221 aggaactgcg ccagttcctc cggatcggtg aagccggaga gatccagcgg ggtctcctcg
     5281 aacacctcga agtcgtgcag gaaggtgaag gcgagcagtt cgcgggcgaa gtcctcggtc
     5341 cgcttccact gcgccccgtc gagcagcgcg gccaggatct cgcggtcgcc ccggaaggcg
     5401 ttgagatgca gttgcaccag gctgtagcgg gagtctcccg catagacgtc ggtgaagtcg
     5461 acgatcccgg tgacctcggt cgcggccagg tccacgaaga tgttggtccc gtgcaggtcg
     5521 ccgtggacga accggggttc gcggccggcc agcagcgtgt ccacgtccgg cagccagtcc
     5581 tccaggcggt ccagcagccg gggcgagagg tagccccacc cgcggtggtc ctcgacggtc
     5641 gccgcgcggc gttcccgcag cagttccggg aagacctcgg aatggggggt gagcacggtg
     5701 ttcccggtca gcggcaccct gtgcagccgg ccgagcaccc ggccgagttc gcgggccagg
     5761 gcgagcagcg cgttccggtc ggtcgtgccg tccatcgcgg accgccaggt ggtgccggtc
     5821 atccggctca tcaccaggta gggccacggc caggctccgg tgccgggccg cagctcgccg
     5881 cggccgagga ggcggggcac cggcaccggg gcgtccgcca ggaccgcgta cgcctccgac
     5941 tccgacgcga ggctctccgg accgcaccag tgctcgccga acagcttgat caccgggccg
     6001 ggctcgccga ccagtacggg gttggtgctc tcgccgggca cccgcagcac cggcggcacc
     6061 ggcagcccga gctcctccag ggctcggcgg gccagcggct cccagaattc ctggtcgttc
     6121 cgcaggctcg cgtaggaatc atccgaatca atacggtcga gaagtaacag ggattcttgt
     6181 gtcacagcgg acctctattc acagggtacg ggccggctta attccgcacg gccggtcgcg
     6241 acacggcctg tccgcaccgc ggatcaggcg ttgacgatga cgggctggtc ggccacgtcg
     6301 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     6361 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     6421 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6481 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatca
//
